Test for Phosphatase enzyme in Milk to determine complete pasteurization of milk
Phosphatase Test on Milk:-
Principle: – Phosphatase is an enzyme present in raw milk which is destroyed by adequate pasteurization, thus one can determine whether the milk has been properly pasteurized by testing for the absence of this enzyme. The milk which in its raw conditions when added to a substrate, disodium phenyl phosphate releases phenol which can be conveniently detected by colorimetric reaction with a suitable reagent.

Disodium phenyl phosphate + Phosphates —> Phosphate +Phenol+Reagent

Procedure:-Mix 0.5ml of milk with 10ml of sodium veronaldisodium phenyl phosphate buffer solution and incubate at 370c for 10min. Add 4 drops of 2,6 dibromo quinine chlorimide solution and keep it for colour development, allow the colour to develop. After 5min. shake with 3 ml. of n-butyl alcohol. Blue colour in n-butyl alcohol indicates presence of phosphates and hence improperly pasteurized milk. A control test was performed on sample of pasteurized milk and the test sample was compared with it. A faint blue colour appeared in pasteurized milk which was used for the comparison test for pasteurization of milk
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